Key metalloproteinases are expressed by specific cell types in experimental autoimmune encephalomyelitis
Key metalloproteinases are expressed by specific cell types in experimental autoimmune encephalomyelitis
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DOI：
10.4049/jimmunol.173.8.5209
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发表时间：
2004-10-15
影响因子：
4.4
通讯作者：
Owens, T
中科院分区：
文献类型：
--
作者：
Toft-Hansen, H;Nuttall, RK;Owens, T
金属蛋白酶(MPS)包括基质金属蛋白酶(MMPs)和金属蛋白酶去整合素(ADAMS)。它们的生理抑制物是金属蛋白酶组织抑制物(TIMPs)。在多发性硬化症及其动物模型实验性自身免疫性脑脊髓炎(EAE)的发病机制中，MPS被认为是细胞浸润的介质。我们使用实时RT-PCR分析了SJL/J小鼠和过继转移髓鞘碱性蛋白(MBP)特异性EAE小鼠脊髓中所有已知的22个小鼠MMPs、7个ADAMS和所有4个已知TIMP的表达。MMP8、MMP10、MMP12、ADAM-12和TIMP-1的表达上调，而MMP15的表达下调。表达水平与病程相关，除ADAM-12外，其余均在缓解期恢复到对照水平。为了研究炎症的中枢神经系统中这些强影响蛋白的潜在细胞来源，我们通过细胞分离从EAE小鼠的中枢神经系统分离出巨噬细胞、粒细胞、小胶质细胞和T细胞，并用实时RT-PCR分析它们的表达。这表明巨噬细胞是基质金属蛋白酶-12和基质金属蛋白酶-1的主要来源。粒细胞是基质金属蛋白酶-8的主要来源。ADAM-12主要由T细胞表达。免疫组织化学染色或原位杂交法证实MMP10、TIMP-1和ADAM-12在血管周围浸润液中的细胞定位。对照组小胶质细胞表达强烈的基质金属蛋白酶-15信号。免疫组织化学染色证实，EAE组小胶质细胞表达的基质金属蛋白酶-15显著下调。我们的研究确定了中枢神经系统炎症中关键MPS的细胞来源。
Metalloproteinases (MPs) include matrix metalloproteinases (MMPs) and metalloproteinase-disintegrins (ADAMs). Their physiological inhibitors are tissue inhibitor of metalloproteinases (TIMPs). MPs are thought to be mediators of cellular infiltration in the pathogenesis of multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). We used real-time RT-PCR to profile the expression of all 22 known mouse MMPs, seven ADAMS, and all four known TIMPs in spinal cord from SJL/J mice and mice with adoptively transferred myelin basic protein (MBP)-specific EAE. A significant and >3-fold alteration in expression was observed for MMP-8, MMP-10, MMP-12, ADAM-12, and TIMP-1, which were up-regulated, and for MMP-15, which was down-regulated. Expression levels correlated with disease course, with all but ADAM-12 returning toward control levels in remission. To examine potential cellular sources of these strongly affected proteins in the inflamed CNS, we isolated macrophages, granulocytes, microglia, and T cells by cell sorting from the CNS of mice with EAE and analyzed their expression by real-time RT-PCR. This identified macrophages as a major source of MMP-12 and TIMP-1. Granulocytes were a major source of MMP-8. ADAM-12 was expressed primarily by T cells. Cellular localization of MMP-10, TIMP-1, and ADAM-12 in perivascular infiltrates was confirmed by immunostaining or in situ hybridization. Microglia from control mice expressed strong signal for MMP-15. Strikingly, the expression of MMP-15 by microglia was significantly down-regulated in EAE, which was confirmed by immunostaining. Our study identifies the cellular sources of key MPs in CNS inflammation.